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基质金属蛋白酶-2,9及其抑制因子-1,2在晶状体上皮细胞的表达及意义 |
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| 基质金属蛋白酶-2,9及其抑制因子-1,2在晶状体上皮细胞的表达及意义 |
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作者：谢二娟 张… 文章来源：华中科技大学同济医学院附属同济医院眼科 点击数：2389 更新时间：2005/4/26 23:47:16 
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| 摘要
目的：研究基质金属蛋白酶-2,9（MMP-2、MMP-9）及其抑制因子-1,2（TIMP-1、TIMP-2）在老年性白内障及透明晶状体上皮细胞和过氧化氢(H2O2)诱导大鼠白内障模型中晶状体上皮细胞的表达，并探讨其在老年性白内障发病中的作用。 方法：随机收集40例老年性白内障患者（排除代谢性和并发性白内障）和8例透明晶状体前囊膜，用免疫组织化学染色(SP-DAB法)和免疫荧光技术对MMP -2、MMP-9、TIMP-1、TIMP-2的表达进行定性、定位检测，并采用HPIAS-1000型医学彩色图像分析系统对结果进行半定量分析。大鼠晶状体随机分为实验组和对照组，实验组使用H2O2诱导，对照组不加H2O2，每组按不同时间点（0、6、12、24、48、72 h）共设6个亚组，每个亚组5只晶状体。用免疫组织化学方法(SP-DAB法)和图像分析系统检测MMP-2、MMP-9、TIMP-1、TIMP-2在大鼠晶状体上皮细胞中的表达。应用SPSS10.0统计软件进行统计学分析。 结果：MMP-2、MMP-9、TIMP-1、TIMP-2阳性表达均位于晶状体上皮细胞的细胞质，胞质内可见程度不等的棕黄色颗粒或绿色荧光。在40例老年性白内障患者，四者的表达均为阳性，而在透明晶状体组8例患者，只有1例四者表达为弱阳性，其余7例均为阴性表达。MMP-2、MMP-9、TIMP-2、TIMP-1在老年性白内障组和透明晶状体组平均吸光度(A值)分别为0.4479±0.0937、0.1458±0.0536；0.5734±0.1094、0.1357±0.0231；0.1779±0.0655、0.0392±0.0093；0.2790±0.0626、0.0922±0.0282；组间差异均有显著性意义（P=0.017，P=0.000，P=0.000，P=0.000）。老年性白内障患者晶状体前囊膜MMP-2和TIMP-2的表达呈显著正相关（r=0.736，P=0.000），MMP-9和TIMP-1的表达呈显著正相关（r=0.627，P=0.000）；MMP-2、MMP-9、TIMP-2、TIMP-1的表达与年龄均无明显相关（r=－0.044、P=0.790，r=－0.080、P=0.624，r=－0.040、P=0.805，r=0.028、P=0.865）。H2O2诱导大鼠白内障模型中，对照组各亚组大鼠晶状体在培养的各个时间段，MMP-2、MMP-9、TIMP-1、TIMP-2表达稳定，均呈弱表达，表达水平均无显著性差异（P >0.05）。和对照组相比，实验组晶状体上皮细胞内四者表达明显增强，差异有显著性意义（P <0.05），而且随着H2O2作用时间延长，各亚组之间四者表达水平呈时间依赖性。在H2O2作用6 h，MMP-2表达开始增高，12 h继续增高，48 h达到高峰，72 h下降。TIMP-2于12 h表达开始增高，48 h达高峰，72 h表达下降。在H2O2作用6 h，MMP-9表达开始增高，48 h达到高峰，以后逐渐下降。TIMP-1于12 h表达开始增高，48 h达高峰，以后逐渐下降。实验组大鼠晶状体前囊膜MMP-2和TIMP-2的表达呈显著正相关（r=0.915，P=0.029），MMP-9和TIMP-1的表达呈显著正相关（r=0.991，P=0.001）。 结论：老年性白内障患者晶状体上皮细胞阳性表达MMP-2、MMP-9、TIMP-2、TIMP-1，表达产物定位于细胞质。而四者在透明晶状体上皮细胞几乎不表达。在H2O2诱导的氧化损伤白内障模型的不同时间段，大鼠晶状体上皮细胞中MMP-2、MMP-9、TIMP-1、TIMP-2的表达发生改变。提示基质金属蛋白酶及其抑制因子可能在老年性白内障发病中发挥重要作用。
Expression of matrix metalloproteinase-2、-9 and their tissue inhibitors -1、-2
on lens epithelial cells
Abstract Objective To study the expression of MMP-2、MMP-9、TIMP-1、TIMP-2 on lens epithelial cells(LECs) of age-related cataract and transparent lens and rat LECs of hydrogen peroxide(H2O2)-induced cataract, and investigate their roles on age-related cataract. Methods Forty cases patients anterior lens capsules of age-related cataract and eight cases normal lens were collected, and were stained by immunohistochemistry and immunofluorescence technique. The results of staining image were analyzed by computer medical image analysis system. Rat lens were cultured in vitro and individed randomly into two groups: experimental group induced by 2 mmol/L H2O2 and control group. There were six subgroups with 5 lens at different time points (0、6、12、24、48、72 h) in each group. The lens were observed under microscope. The expression of MMP-2、MMP-9、TIMP-1、TIMP-2 was detected with immunohistochemical staining methods, and the results(A) were analyzed by computer medical image analysis system. Results In forty cases age-related cataract patients group, the expression of MMP-2、MMP-9、TIMP-2、TIMP-1 were all positive. However, eight cases normal lens, seven negative and only one positive. There was significant difference of expression of MMP-2、MMP-9、TIMP-2 and TIMP-1 between age-related cataract LECs and normal LECs(P=0.017，P=0.000，P=0.000，P=0.000). In the age-related group, the expression of MMP-2 and TIMP-2 presented a linear positive correlation（r=0.736，P=0.000）, and the expression of MMP-9 and TIMP-1 presented a linear positive correlation also（r=0.627，P=0.000）. There were no linear correlations between the expression of MMP-2、MMP-9、TIMP-2、TIMP-1 and the age of age-related patients(r=－0.044、P=0.790，r=－0.080、P=0.624，r=－0.040、P=0.805，r=0.028、P=0.865). In rat H2O2-induced cataract model, the expression of MMP-2、MMP-9、TIMP-1、TIMP-2 was stable and weak in the control group, and there were no significant difference among all subgroups. When the lens were cultured with 2 mmol/L H2O2 for 6 h, the expression of MMP-2 increased obviously, and peaked at 48 h, decreased at 72 h. TIMP-2 expression elevated at 12 h, peaked at 48 h, and then dropped gradually. The expression of MMP-9 increased at 6 h, reached peaks at 48 h, and decreased gradually. TIMP-1 expression increased at 12 h, reached peaks at 48 h, and decreased at 72 h. In the H2O2 group, the expression of MMP-2 and TIMP-2 presented a linear positive correlation(r=0.915，P=0.029), and the expression of MMP-9 and TIMP-1 presented a linear positive correlation(r=0.991，P=0.001). Conclusion The expression of MMP-2、MMP-9、TIMP-2、TIMP-1 on lens epithelial cells of age-related cataract is positive, and those of transparent lens are negative. H2O2 can upregulate the expression of MMP-2、MMP-9、TIMP-1、TIMP-2 on rat LECs. It indicates that they may play an important role in the formation of age-related cataract.
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| 会议投稿录入：一刀_2002 责任编辑：毛进 |
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